*Target (protein/gene namephosphatidylinositol 4-kinase, putative (Plasmodium vivax ) *NCBI Gene # or RefSeq#:126793/ XM_001613220.1. *Protein ID (NP or XP #) or Wolbachia#:XP_001613270.1
*Organism (including strain):Plasmodium vivax Etiologic Risk Group (see link below): */Disease Information (sort of like the Intro to your Mini Research Write up): Plasmodium vivax is both a pathogenic protozoal parasite. It one of the five species is responsible for human malaria. Although not as deadly as Plasmodium falciparum. It can cause sever disease and death via splenomegaly. They are most commonly found in female Anopheles mosquito and only be transmitted by female because only female mosquito bites. Plasmodium vivax originated in the United States, Latin America, and small part of Africa; however, it has now an emerging problem for developing centuries. Once red blood cells of hosts are infected by Plasmodium vivax, they rupture, leading to fever. Ruptured red blood cell also have tendency to stick to endothelia of blood vessels. When vessels are clogged, RBC deliver oxygen to tissues efficiently causing oxygen deprivation of the tissue. Some other symptoms may also include enlargement of the spleen. Additionally, the protozoal parasite can go dormant for days to years. The primary treatment for Plasmodium vivax is chloroquine.
Link to TDR Targets page (if present): N/A
Link to Gene Database page (NCBI, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.)
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3940870/ - The Plasmodium vivax phosphatidylinositol 4-kinase (PI4K) is an essential protein required for the life stage of PI4KIIIβ. PI4KIIIβ is a ubiquitous eukaryotic enzyme that phosphorylates lipids to regulate intracellular signaling and trafficking, hence often a target for drug discovery. To be more specific, Type III phosphatidylinositol 4-kinase (PI4KIIIbeta) is an essential enzyme for organizing membrane trafficking, which is required for a variety of pathogenic processes.
Purification Method X-RAY DIFFRACTION Image of protein (PyMol with features delineated and shown separately):
Figure 2. PyMOL cartoon representation of Phosphatidylinositol 4-kinase III beta-PIK93 in a complex with Rab11a- GTP gammaS found in E. Coli; PDB file 4D0L
*Amino Acid Sequence (paste as text only - not as screenshot or as 'code'):
>EDL43543.1 phosphatidylinositol 4-kinase, putative [Plasmodium vivax]
MDHKTDGKEDHKREGKTEGETGQTGKEDAGDTTEGADQREKWTAAPEESTSYRSLYKHNYKAHVYADDFV
CLRLKGRGGEGHDDGSSEPQQCVEPFEGDLPVGGCPHDDTPVGELSTKRDNNEEVAATGRDNLCGVAVDR
IDEANPPREDAQYVQVGSRMEDHQPDEAIYSQAGTQHSNHANEEKRENSDLQPHSDDNYTLQESYSSDCH
LKEEMPHESNQPSEDLPSWEAHKMKEKDQPPEKQNKIKIIEMFKMKLESKEMNRSRHFGDDVRGGGPGGA
HHDGYGDGRDDGRDDGRDDGHDDGHGDGHDHSGGASPGASNRCTQIRESISRIFSESPSSKPKIISELKH
GEGHPNSGSNGSGSHHSSGTHHGSGSNGSGSHERERADGKANGAKNALKEGSLLRLFRCEYFDTHLHIRY
LYDRREVGVHEYLVNSLYTQRKYEDILFYLPQLSQISLVRYESSSLYRFLLCKASNSMHFALKLSWIYHS
IVEDNSSKYKDLAHKMTQEIEMAVVNCKPFNGKGTSGNENKQSYLLNLAHPLLFKRKYLIKRIKDNERFE
RTKLSRNCLNRSCNLCLLKGGEESTQSGMEVAVQGETQRNSQSADLSSAPNAAPSAAPNAAAHAATHAAA
HASPKKRRKREATSTAPPPKLPQCYIVNSGALSIARARVKLPSTYAKLGNPLSASKFFLPEFNCTFDMIG
ELQQFFMKQRRCDYFSLLNNFVNLTISVSNLLSTEPDVEIRNELLNRFIYSLNSWMLMRRCIVAACTNVF
SMTGLCIPLECLSPPQSGQRTEPTPSSNGSNSNGSSNISGSSGSSGSGLQILHFNYDECKIFFSKKRAPY
LLMFEVADLDEDISHIPDGLFYPSSVGGDEAAGEEEEEAGQTASETAGEAADGATRGVTDEPTSQPTPPP
RRKGHRIERNFCSSKNYGLFNENSSSYDYEPRGGVHDGKRKKKKKKKKRRSESGEDDEGGAHTGQSGSEV
DRGKEERGKGAQGKKTTGADLDALKMEHMYVYNCIVNDLRKENLISFPSGEEDSLSIIRKCIGMRADEED
EEGGDTEGGDTNGGDPGDPSEPSNLGQGGPGWGAPPAEEGPREATKSFLVNARSASMPNYLSSSLERCNI
SSNDNDSNEVKTSVESISSGLADLPPGGLHGSEVDAQVDAQEDAQVEAPDGGWSPSPGKRPNDSGVTADQ
PLEGAAPSPHSSFLHAGEKTGDALDIDEYFKPANYTNEEFKKRYCRVIKRLLWGELFEEKKKKIRKTSAY
GKLKTWDLKCVIVKGGDDLRQELLASQLIRQFKIIFENAGLPLWLRPYEILVTGANSGIIEYVNDTCSVD
SLKRKFGADSISTIFNVVFADYIFEAKKNFIESHAAYSLVSYLLQVKDRHNGNLLLDSDGHLIHIDYGFM
LTNSPGNVNFETSPFKLTQEYLDIMDGEKSENYEYFRRLIVSGFLEARKHSEEIILFVELMMPALKIPCF
ANGTQFCIDSLKERFMTNLAVDVCIQRINALIESSINNFRSVQYDYFQRITNGIM
*length of your protein in Amino Acids 1525 aa
Molecular Weight of your protein in kiloDaltons using the Expasy ProtParam website
168828.92 Da
Molar Extinction coefficient of your protein at 280 nm wavelength:125875 mol-1 cm-1
)
*NCBI Gene # or RefSeq#: 126793/ XM_001613220.1.
*Protein ID (NP or XP #) or Wolbachia#:XP_001613270.1
*Organism (including strain): Plasmodium vivax
Etiologic Risk Group (see link below):
*/Disease Information (sort of like the Intro to your Mini Research Write up):
Plasmodium vivax is both a pathogenic protozoal parasite. It one of the five species is responsible for human malaria. Although not as deadly as Plasmodium falciparum. It can cause sever disease and death via splenomegaly. They are most commonly found in female Anopheles mosquito and only be transmitted by female because only female mosquito bites. Plasmodium vivax originated in the United States, Latin America, and small part of Africa; however, it has now an emerging problem for developing centuries. Once red blood cells of hosts are infected by Plasmodium vivax, they rupture, leading to fever. Ruptured red blood cell also have tendency to stick to endothelia of blood vessels. When vessels are clogged, RBC deliver oxygen to tissues efficiently causing oxygen deprivation of the tissue. Some other symptoms may also include enlargement of the spleen. Additionally, the protozoal parasite can go dormant for days to years. The primary treatment for Plasmodium vivax is chloroquine.
Link to TDR Targets page (if present): N/A
Link to Gene Database page (NCBI, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.)
https://www.ncbi.nlm.nih.gov/gene/?term=phosphatidylinositol+4-kinase%2C+putative%C2%A0Plasmodium+vivax
Essentiality of this protein:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3940870/ -
The Plasmodium vivax phosphatidylinositol 4-kinase (PI4K) is an essential protein required for the life stage of PI4KIIIβ. PI4KIIIβ is a ubiquitous eukaryotic enzyme that phosphorylates lipids to regulate intracellular signaling and trafficking, hence often a target for drug discovery. To be more specific, Type III phosphatidylinositol 4-kinase (PI4KIIIbeta) is an essential enzyme for organizing membrane trafficking, which is required for a variety of pathogenic processes.
Is it a monomer or multimer as biological unit? (make prediction athttp://www.ebi.ac.uk/msd-srv/prot_int/pistart.html): Monomer
Complex of proteins?:
Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism):
http://pubs.acs.org/doi/ipdf/10.1021/acs.jmedchem.5b01311
*EC#: 2.7.1.67
Link to BRENDA EC# page:
http://www.brenda-enzymes.org/enzyme.php?ecno=2.7.1.67
-- Show screenshot of BRENDA enzyme mechanism schematic
Figure 1. BRENDA enzyme mechanism schematic of the 1-phosphatidylinositol 4 kinase ( 2.7.1.67).
Enzyme Assay information (spectrophotometric, coupled assay ?, reagents):
http://www.jbc.org/content/277/22/20041.full
-- link to Sigma (or other company) page for assay (see Sigma links below)
-- -or link (or citation) to paper that contains assay information
-- links to assay reagents (substrates) pages.
--- List cost and quantity of substrate reagents, supplier, and catalog #
1 g ATP (Sigma, A2383) - $52.50
Structure (PDB or Homology model)
-- For Homology Model option:
---- Show pairwise alignment of your BLASTP search in NCBI against the PDB
---- Query Coverage:23%
---- Max % Identities:44%
---- % Positives 67%
---- Chain used for homology: Chain A, Phosphatidylinositol 4-kinase Iii Beta-pik93 In A Complex With Rab11a-gtp Gammas
Current Inhibitors: imidazopyrazines
Expression Information (has it been expressed in bacterial cells): Has been expressed in E. coli
https://www.ncbi.nlm.nih.gov/pubmed/?term=24876499
Purification Method X-RAY DIFFRACTION
Image of protein (PyMol with features delineated and shown separately):
Figure 2. PyMOL cartoon representation of Phosphatidylinositol 4-kinase III beta-PIK93 in a complex with Rab11a- GTP gammaS found in E. Coli; PDB file 4D0L
*Amino Acid Sequence (paste as text only - not as screenshot or as 'code'):
>EDL43543.1 phosphatidylinositol 4-kinase, putative [Plasmodium vivax]
MDHKTDGKEDHKREGKTEGETGQTGKEDAGDTTEGADQREKWTAAPEESTSYRSLYKHNYKAHVYADDFV
CLRLKGRGGEGHDDGSSEPQQCVEPFEGDLPVGGCPHDDTPVGELSTKRDNNEEVAATGRDNLCGVAVDR
IDEANPPREDAQYVQVGSRMEDHQPDEAIYSQAGTQHSNHANEEKRENSDLQPHSDDNYTLQESYSSDCH
LKEEMPHESNQPSEDLPSWEAHKMKEKDQPPEKQNKIKIIEMFKMKLESKEMNRSRHFGDDVRGGGPGGA
HHDGYGDGRDDGRDDGRDDGHDDGHGDGHDHSGGASPGASNRCTQIRESISRIFSESPSSKPKIISELKH
GEGHPNSGSNGSGSHHSSGTHHGSGSNGSGSHERERADGKANGAKNALKEGSLLRLFRCEYFDTHLHIRY
LYDRREVGVHEYLVNSLYTQRKYEDILFYLPQLSQISLVRYESSSLYRFLLCKASNSMHFALKLSWIYHS
IVEDNSSKYKDLAHKMTQEIEMAVVNCKPFNGKGTSGNENKQSYLLNLAHPLLFKRKYLIKRIKDNERFE
RTKLSRNCLNRSCNLCLLKGGEESTQSGMEVAVQGETQRNSQSADLSSAPNAAPSAAPNAAAHAATHAAA
HASPKKRRKREATSTAPPPKLPQCYIVNSGALSIARARVKLPSTYAKLGNPLSASKFFLPEFNCTFDMIG
ELQQFFMKQRRCDYFSLLNNFVNLTISVSNLLSTEPDVEIRNELLNRFIYSLNSWMLMRRCIVAACTNVF
SMTGLCIPLECLSPPQSGQRTEPTPSSNGSNSNGSSNISGSSGSSGSGLQILHFNYDECKIFFSKKRAPY
LLMFEVADLDEDISHIPDGLFYPSSVGGDEAAGEEEEEAGQTASETAGEAADGATRGVTDEPTSQPTPPP
RRKGHRIERNFCSSKNYGLFNENSSSYDYEPRGGVHDGKRKKKKKKKKRRSESGEDDEGGAHTGQSGSEV
DRGKEERGKGAQGKKTTGADLDALKMEHMYVYNCIVNDLRKENLISFPSGEEDSLSIIRKCIGMRADEED
EEGGDTEGGDTNGGDPGDPSEPSNLGQGGPGWGAPPAEEGPREATKSFLVNARSASMPNYLSSSLERCNI
SSNDNDSNEVKTSVESISSGLADLPPGGLHGSEVDAQVDAQEDAQVEAPDGGWSPSPGKRPNDSGVTADQ
PLEGAAPSPHSSFLHAGEKTGDALDIDEYFKPANYTNEEFKKRYCRVIKRLLWGELFEEKKKKIRKTSAY
GKLKTWDLKCVIVKGGDDLRQELLASQLIRQFKIIFENAGLPLWLRPYEILVTGANSGIIEYVNDTCSVD
SLKRKFGADSISTIFNVVFADYIFEAKKNFIESHAAYSLVSYLLQVKDRHNGNLLLDSDGHLIHIDYGFM
LTNSPGNVNFETSPFKLTQEYLDIMDGEKSENYEYFRRLIVSGFLEARKHSEEIILFVELMMPALKIPCF
ANGTQFCIDSLKERFMTNLAVDVCIQRINALIESSINNFRSVQYDYFQRITNGIM
*length of your protein in Amino Acids
1525 aa
Molecular Weight of your protein in kiloDaltons using the Expasy ProtParam website
168828.92 Da
Molar Extinction coefficient of your protein at 280 nm wavelength: 125875 mol-1 cm-1
TMpred graph Image(http://www.ch.embnet.org/software/TMPRED_form.html). Input your amino acid sequence to it.
*CDS Gene Sequence (paste as text only):
atggaccaca aaacagatgg caaagaagac cacaaaagag aaggcaaaac tgaaggcgaa
acgggacaga ccggcaagga agatgctggc gacacaaccg agggcgccga ccaaagggaa
aagtggaccg ccgcaccgga agagagcaca tcgtataggt ctctgtacaa acacaactac
aaagcgcacg tgtacgcgga tgattttgtg tgccttcgat tgaaaggtcg tggaggtgag
gggcacgatg atggaagcag tgaaccgcag cagtgtgtag agccgttcga gggggatctc
ccagtgggtg gctgcccaca tgatgataca cctgttgggg agttaagtac taaaagggac
aacaatgagg aggttgctgc cacagggagg gacaacctgt gtggggttgc agtcgatagg
attgacgaag caaacccccc aagggaggat gctcagtacg ttcaggtagg cagcagaatg
gaggaccacc aaccagacga agcgatatac tcccaagcgg ggacacaaca tagcaatcac
gctaatgagg agaagaggga aaactcggat cttcagcctc actccgatga taattacacc
cttcaagaaa gctacagcag tgactgccat ctgaaggagg aaatgccaca cgagagtaac
caacccagtg aggacctgcc cagctgggaa gcacacaaaa tgaaggagaa ggaccaaccc
ccagagaaac aaaacaaaat taaaattatt gaaatgttta agatgaagct ggagagtaag
gagatgaaca ggagtcgcca tttcggtgat gacgtccgtg gggggggtcc cggtggcgct
catcacgatg ggtatggcga tgggcgtgac gatgggcgtg acgatgggcg tgacgatggg
catgacgatg ggcatggtga tgggcatgac cactcggggg gcgcctcccc cggtgcctct
aaccgctgca cgcagatacg ggagagcatc tctcgcatat tcagcgagtc gccctcctcc
aagccgaaga taataagcga gctgaagcac ggcgaggggc accccaacag cggtagcaac
ggaagcggca gccaccacag cagcggtacc caccacggaa gcggtagcaa cggaagcggc
agccacgaac gggagagggc ggacggaaag gccaacggag cgaagaacgc attgaaggag
ggaagcctct tgaggctctt cagatgcgaa tactttgaca cccacctgca cattaggtac
ctctacgaca ggagagaagt gggggtccac gagtacctgg tgaattccct ttacacgcag
aggaagtacg aggatatatt attttacctc ccgcagctca gtcaaatttc gctagtccgt
tacgagtcct cttccctcta tcgcttcctc ctttgcaaag ccagcaactc catgcacttc
gcattaaagc tcagctggat ctatcattct atagtagagg acaactcttc aaaatataag
gacttggctc acaagatgac tcaagaaatt gaaatggctg tggtgaattg caaaccattt
aacggtaaag gcacctcggg gaacgagaat aagcagtcgt acctgctgaa cctcgcccac
ccgctcctct tcaaaaggaa gtacctcatt aaaaggatca aagataatga gcggttcgag
cgcacgaagc tatccaggaa ttgcctgaac aggtcatgca acttgtgctt gctgaagggg
ggggaagagt ccacgcagag cgggatggaa gtggctgtgc aaggcgaaac gcagaggaac
tcccaaagcg ctgatctgag ttccgctcca aatgccgctc caagcgccgc tccaaacgcc
gctgctcatg ccgccacaca tgccgctgca catgcctcgc ccaagaagcg gcgcaaacgg
gaggcgacga gcacagcgcc gccccccaag ctgccccagt gctacattgt caactcgggc
gccctctcca tagccagggc cagagtgaag ctccccagca cctacgccaa gctgggcaac
cccctcagcg catccaagtt cttcctccca gaatttaatt gcacctttga tatgatagga
gagctgcagc agttttttat gaagcagagg aggtgcgatt actttagcct cctgaacaac
tttgtgaact tgaccatctc cgtttctaac cttctgtcca ccgaaccaga tgtagagata
cgcaacgagc tcctcaacag attcatttac tctttaaaca gttggatgct catgcgtagg
tgcatcgtgg ctgcatgtac taacgttttt tccatgacgg ggctgtgcat tccgctcgag
tgcctgtccc ccccccagag cggccagcga acggagccga ccccaagcag caatggcagc
aatagcaacg gtagcagcaa catcagcggt agcagcggca gcagcggcag cggcttgcag
atccttcact ttaactacga tgagtgcaaa atatttttct ccaagaagag ggcgccctac
ctgctcatgt tcgaggtggc ggacctggac gaggacatca gccacattcc ggacggtctc
ttttacccct ccagcgtggg gggggatgaa gcggcgggtg aggaggagga agaagcaggt
caaacggcga gcgaaacggc gggtgaggca gcggatggtg caacccgtgg cgtgactgac
gaaccgacgt cgcaaccgac gccgccaccg cgaaggaagg gccaccgcat cgaaaggaac
ttctgttcct ctaaaaacta cgggctcttc aacgagaaca gcagctccta cgactacgag
ccgcgaggcg gggtgcatga tgggaagagg aagaagaaga agaaaaagaa gaagaggaga
agcgaaagtg gggaggacga cgaggggggg gcgcacaccg gccagagtgg ttcagaagtg
gatcggggga aggaggaacg cgggaaaggc gcgcagggaa aaaaaacaac cggcgcggac
ctggacgctc tgaaaatgga gcacatgtac gtatacaact gcatagtgaa cgacctgcga
aaggagaatt tgatttcgtt cccctcgggg gaggaggaca gcctctccat aattaggaag
tgcataggga tgagggcgga cgaggaggac gaggaggggg gagacacgga ggggggcgac
acgaacgggg gcgatccggg cgatccgagc gaaccgagca acctcgggca gggcggcccc
ggttgggggg ccccccccgc ggaggaaggc ccccgggagg caaccaagag cttcctggtg
aacgcccgct ccgcgtctat gcccaactat ctaagtagct ccctggagag gtgcaacatc
agctcgaacg acaatgacag caatgaagtg aaaacgtctg tggagagcat cagcagtggg
ctggctgacc tcccacctgg ggggttgcac ggaagtgagg tggacgctca ggtggacgcc
caggaggatg cccaggtgga agctccagat ggaggatgga gcccctcccc gggtaaacgc
cccaatgata gtggtgtcac tgcagatcaa ccgttagaag gcgccgcccc ctccccacac
agcagcttct tacacgcggg ggagaaaaca ggcgacgcgc tagacatcga cgagtacttc
aaaccagcga attacaccaa cgaggaattt aaaaaaaggt attgtagagt tataaagagg
ctgctatggg gagagctatt tgaagaaaaa aaaaaaaaaa ttaggaaaac gtcagcttat
gggaagttaa agacatggga tttgaagtgc gtaatcgtta agggtgggga tgacttgagg
caggagctgc tggcatccca actgattcgc cagtttaaaa ttatttttga gaatgcgggc
ctgccgctgt ggctacgccc ctacgaaatt ctagtgacgg gggcaaactc cggcatcatt
gaatacgtaa atgacacctg ctcggtggac tcgctcaagc ggaagttcgg cgccgacagc
atctccacca tcttcaacgt ggtcttcgcg gactacatct tcgaggccaa gaagaacttc
atcgagagcc acgcagccta ctcgctcgtc tcgtacctcc tccaggtgaa ggacaggcac
aacggaaacc tcctgctgga ctcggacgga cacctcatcc acatagacta cggcttcatg
ctcaccaact cccctggaaa cgtaaatttt gagacctccc ccttcaaact cacccaagaa
tacctagaca tcatggacgg agaaaagtca gaaaattatg agtacttcag gcgcctcatc
gtcagtggct tcctcgaggc ccgcaagcac tcggaggaga tcatcctctt cgtcgagctg
atgatgccag ctttaaaaat cccctgcttt gcgaacggca cgcagttctg catcgactcg ctgaaggagc gcttcatgac gaacctggcc gtcgacgtgt gcatccaacg aatcaacgcc
ctcattgagt cctccatcaa taattttaga agcgtccagt acgactactt ccagagaatc
accaacggaa ttatgtga
*GC% Content for gene: 54.9%
*CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only):
*GC% Content for gene (codon optimized):